miR-342-3p对鼻咽癌细胞增殖、凋亡的影响及可能机制

Effect and possible mechanism of miR-342-3p on the proliferation and apoptosis of nasopharyngeal carcinoma cells

目的探讨Aβ1-42对小鼠巨噬细胞RAW264.7向炎症性细胞转变的影响及机制。方法采用实时荧光定量PCR(qRT-PCR)检测鼻咽上皮细胞NP69和鼻咽癌细胞CNE-1、CNE-2、HNE-1、5-8F、HONE-1中miR-342-3p的表达水平。采用脂质体介导法向CNE-2细胞转染miR-342-3p模拟物记为miR-342-3p mimic组,转染模拟物对照的CNE-2细胞记为NC组,不行转染的CNE-2细胞记为Control组。以qRT-PCR检测转染效果,四甲基偶氮唑盐(MTT)法和克隆形成实验检测转染对细胞增殖的影响,Annexin-Ⅴ-FITC/PI双染色法检测转染对细胞凋亡的影响,Western blot检测细胞中caspase-3、Ki-67及NF-κB信号通路相关蛋白表达水平。结果与鼻咽上皮细胞NP69相比,miR-342-3p在鼻咽癌细胞中的表达水平显著降低。与Control组和NC组相比,miR-342-3p mimic组细胞miR-342-3p的表达水平显著升高,细胞OD值明显降低,克隆形成率显著下降,凋亡显著升高(P<0.05),细胞中caspase-3激活水平升高,Ki-67、p-IKKα、p-IKKβ蛋白水平降低(P<0.05)。结论 miR-342-3p在鼻咽癌细胞中呈低表达,上调鼻咽癌CNE-2细胞中miR-342-3p的表达能够抑制细胞增殖,促进细胞凋亡,其作用与抑制NF-κB信号通路的激活相关。

Objective To investigate the expression of miR-342-3 p in nasopharyngeal carcinoma cells and its effect on proliferation and apoptosis of nasopharyngeal carcinoma cells and possible mechanism.Methods qRT-PCR was used to detect the expression of miR-342-3 p in nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells(CNE-1,CNE-2,HNE-1,5-8 F and HONE-1).The transfection effect was detected by qRT-PCR,the effect of transfection on cell proliferation was detected by MTT assay and colony formation assay,and apoptosis was detected by Annexin-V-FITC/PI double staining.The expression of caspase-3,Ki-67 and NF-κB signaling pathway-related proteins was detected by Western blot.Results The expression level of miR-342-3 p in nasopharyngeal carcinoma cells was significantly lower than that in nasopharyngeal epithelial cells NP69(P<0.05).Compared with the control group and NC group,the expression level of miR-342-3 p in the miR-342-3 p mimic group was increased(P<0.05),the OD value and colony formation rate and levels of Ki-67 and p-IKKαand p-IKKβproteins were decreased(P<0.05),but apoptosis rate and caspase-3 activation level was significantly increased(P<0.05).Results The expression level of miR-342-3 p in nasopharyngeal carcinoma cells was significantly lower than that in nasopharyngeal epithelial cells NP69(P<0.05).Compared with the control group and NC group,the expression level of miR-342-3 p in the miR-342-3 p mimic group was increased(P<0.05),the OD value and colony formation rate and levels of Ki-67 and p-IKKαand p-IKKβproteins were decreased(P<0.05),but apoptosis rate and caspase-3 activation level was significantly increased(P<0.05).Conclusion Up-regulation of miR-342-3 p expression in nasopharyngeal carcinoma CNE-2 cells inhibited cell proliferation and promoted cell apoptosis,which is related to inhibition of NF-activation of theκB signaling pathway.